Cherley Borba Vieira de Andrade

Chemotherapy and radiation regimens to breast cancer treatment induce changes in mRNA levels of renin-angiotensin system related genes in cardiac tissue

Background and aim: Cardiovascular complications are one limitation of breast cancer treatment. The aim of the current study was to investigate whether the renin–angiotensin related genes could be altered by chemotherapy and radiotherapy, using a rat model. Methods: Female rats were divided into three groups: control, chemotherapy + irradiation (TC+IR) and irradiation (IR). Molecular analyses of the left ventricle were performed five months after the end of treatment. The analyses evaluated the changes in mRNA levels of some renin–angiotensin system (RAS) related genes: angiotensinogen, renin, angiotensin-converting enzyme (ACE) and angiotensin II type 1 receptor (AT1) and vascular endothelial growth factor (VEGF), which can be related to ACE production, by RT-PCR. Results: Renin was only observed in treated groups, TC+IR and IR, compared with the control group. ACE and VEGF levels were decreased in TC+IR (p<0.001) and in IR (p<0.001), and AT1 mRNA was higher in groups TC+IR (p<0.01) and IR (p<0.05) compared with the control group. Conclusion: Chemotherapy and irradiation can induce significant changes in some RAS related genes. These alterations are important to understand the pathways and consequences beyond cardiotoxicity induced by breast cancer treatments.

Caspase-3 activation and increased procollagen type I in irradiated hearts

The caspase-3-cleaved presence was evaluated in this study in the heart of irradiated rats, during the decline of ventricular function. Female Wistar rats were irradiated with a single dose of radiation (15 Gy) delivered directly to the heart and the molecular, histological and physiological evaluations were performed at thirteen months post-irradiation. The expressions of procollagen type I, TGF-ß1 and caspase-3-cleaved were analyzed using Western blotting. Cardiac structural and functional alterations were investigated by echocardiography and electron microscopy. In the irradiated group, the levels of procollagen type I, TGF-ß1 and caspase-3-cleaved are increased. Significant histological changes (degeneration of heart tissue and collagen deposition) and functional (reduced ejection fraction) were observed. Data suggest that the cardiac function decline after exposure to ionizing radiation is related, in part, to increased collagen and increased caspase-3-cleaved.

Evaluation of rib microstructure in Wistar rats using SR-μCT after radiation therapy simulation for breast cancer

A better understanding of biological interactions that occur after exposure to photon radiation is needed in order to optimize therapeutic regimens and facilitate development and strategies that decrease radiation-induced side effects in humans. In this work, ribs of Wistar rat submitted to radiotherapy simulation were imaged using synchrotron radiation computed microtomography at Elettra Synchrotron Laboratory in Trieste, Italy. Histomorphometric parameters were calculated directly from the 3D microtomographic images and showed significant differences between irradiated and non-irradiated groups.

Analysis of femur head microstructure in ovariectomized rats

It is well accepted that rat ovariectomy (OVX) is a model of estrogen deficiency. OVX played a very important role in the initiating and developing of osteoporosis and it has been shown to be a major risk factor for the development of postmenopausal osteoporosis in women. In this work we used synchrotron radiation computed microtomography to investigate the skeletal effects in femoral head of female Wistar rats after bilateral ovariectomy surgery. The CT system was set up at the SYRMEP beamline in the synchrotron radiation facility ELETTRA (Trieste, Italy). Micro-CT images provided 3D information on precise trabecular microstructure by the reconstruction of multiple 2D images with almost 2 μm resolution. Our aim was to use histomorphometric analysis to reveal the effect of OVX on the three-dimensional (3D) trabecular bone microarchitecture. Evaluated morphometric parameters were trabecular bone volume-tissue volume ratio (BV/TV), trabecular number (Tb.N), trabecular separation (Tb.Sp) and trabecular thickness (Tb.Th). OVX group presented noticeable reduction in the Tb.N and Tb.Th when compared with control group (P 0.05). Our data may help to gain more insight into the potential mechanism of osteoporotic femoral head fractures.

Evaluation of photon irradiation treatment upon calcium content of ribs of Wistar rats using micro-XRF

Therapeutic doses of radiation have been shown to have deleterious consequences on bone health. Among the treatment strategies used for breast cancer treatment, the most used are radiotherapy and chemotherapy. Radiotherapy may be given to destroy the cancer cells using high-dose x-rays. Protocols vary considerably, but generally whole body irradiation totals from 10 to 15 Gy, whereas local therapy totals from 40 to 70 Gy. In clinical practice, the quantitative evaluation of bone tissue relies on measurements of bone mineral density values, which are closely associated with the risk of osteoporotic fracture. Improved survivorship rates of cancer patients receiving radiotherapy increase the importance of understanding the mechanisms and long-term effects of radiation-induced bone loss. In this work, we investigated the variation on calcium distribution in ribs of female Wistar rats (Rattus norvegicus) submitted to photon irradiation with a single dose of 20 Gy. The determination of the calcium distribution ...

Radiotherapy-Induced Skin Reactions Induce Fibrosis Mediated by TGF-β1 Cytokine

Purpose: This study aimed to investigate radiation-induced lesions on the skin in an experimental animal model. Methods and Materials: Cutaneous wounds were induced in Wistar rats by 4 MeV energy electron beam irradiation, using a dose rate of 240 cGy/min, for 3 different doses (10 Gy, 40 Gy, and 60 Gy). The skin was observed 5, 10, and 25 days (D) after ionizing radiation exposition. Results: Infiltrate inflammatory process was observed in D5 and D10, for the 40 Gy and 60 Gy groups, and a progressive increase of transforming growth factor β1 is associated with this process. It could also be noted a mischaracterization of collagen fibers at the high-dose groups. Conclusion: It was observed that the lesions caused by ionizing radiation in rats were very similar to radiodermatitis in patients under radiotherapy treatment. Advances in Knowledge: This study is important to develop strategies to prevent radiation-induced skin reactions.

Synchrotron radiation X-ray microtomography and histomorphometry for evaluation of chemotherapy effects in trabecular bone structure

Three-dimensional microtomography has the potential to examine complete bones of small laboratory animals with very high resolution in a non-invasive way. One of the side effects caused by some chemotherapy drugs is the induction of amenorrhea, temporary or not, in premenopausal women, with a consequent decrease in estrogen production, which can lead to bone changes. In the present work, the femur heads of rats treated with chemotherapy drugs were evaluated by 3D histomorphometry using synchrotron radiation microcomputed tomography. Control animals were also evaluated for comparison. The 3D tomographic images were obtained at the SYRMEP (SYnchrotron Radiation for MEdical Physics) beamline at the Elettra Synchrotron Laboratory in Trieste, Italy. Results showed significant differences in morphometric parameters measured from the 3D images of femur heads of rats in both analyzed groups.

Analysis of calcium distribution in femur of female rats submitted to different chemotherapy regimens

The most used treatment strategies for breast cancer (BC) are surgery, chemotherapy, and/or radiotherapy. Premenopausal women undergoing adjuvant chemotherapy BC treatment have significant bone loss from the first year after the treatment. This high bone mineral density loss can lead to an increased risk of fractures. Doxorubicin associated with Cyclophosphamide (AC) is a multidrug widely used for BC treatment, although this association can cause severe side effects. Today it is been discussed the use of the Docetaxel and cyclophosphamide (TC) association for BC treatment. The influence on the bone during chemotherapy regimens that include taxanes, like docetaxel, is unknown. Data from the “Women`s Health Initiative” show that postmenopausal BC survivors have a 15% greater risk of developing fractures than women without a history of BC. In this study, it was evaluated parameters involved in osteoporosis when rats were subjected to a chemotherapy regimen (TC) and/or irradiation (IR). Female Wistar rats, 03 months old, were divided into 3 groups: control, TC+IR (G1) and AC+IR (G2). The animals were euthanized 5 after months the end of treatment and their femurs were excised and dissected. Sections of 10 μm thick were used for μXRF analysis at the National Laboratory of Synchrotron Light. The uteri of these rats were collected and weighed Statistical analyzes were performed using GraphPad Prism, and values were compared using ANOVA and Tukey post-hoc test. The obtained results showed that animals from G2 had a significant reduction (p<;0.05) of uterine mass when compared to control. The qualitative analysis performed by μXRF showed that the animals from G2 had iron in bone composition of the femurs. This same result was not observed in animals from G0 and G1 groups. These results suggest that early menopause occurs and osteoporosis begins, probably because of the absence, or reduced, production of estrogen. The presence of iron in the G2 samples indicates the process of osteoporosis, because according to literature, this ion is competitive with calcium ions.

Quantitative computed microtomography for evaluation of radiotherapy effects on bone microstructure

In this work we investigated the consequences of irradiation in the femur of rats to a radiation dose of 5 Gy. Three different sites in femur specimens were imaged using synchrotron radiation computed microtomography to assess trabecular bone microarchitecture: femur head, distal metaphysis and distal epiphysis. Histomorphometric quantification was extracted directly from the 3D tomographic images using synchrontron radiation. The 3D tomographic images were obtained at the SYRMEP (SYnchrotron Radiation for MEdical Physics) beamline at the Elettra Synchrotron Laboratory in Trieste, Italy. A better understanding of the biological interactions that occur after exposure to photon radiation is needed in order to optimize therapeutic regimens and facilitate development and strategies that decrease radiation-induced side effects in humans. Results showed significant differences between irradiated and non-irradiated mostly in head and distal metaphysis bone sites.

Evaluation of Radiation Dose Effects on Rat Bones Using Synchrotron Radiation Computed Microtomography

In this work, we investigated the consequences of irradiation in the femora and ribs of rats submitted to radiation doses of 5 Gy. Three different sites in femur specimens (head, distal metaphysis and distal epiphysis) and one in ribs (ventral) were imaged using synchrotron radiation microcomputed tomography to assess trabecular bone microarchitecture. Histomorphometric quantification was calculated directly from the 3D microtomographic images using synchrotron radiation. The 3D microtomographic images were obtained at the SYRMEP (SYnchrotron Radiation for MEdical Physics) beamline at the Elettra Synchrotron Laboratory in Trieste, Italy. A better understanding of the biological interactions that occur after exposure to photon radiation is needed in order to optimize therapeutic regimens and facilitate development and strategies that decrease radiation‐induced side effects in humans. Results showed significant differences between irradiated and non‐irradiated specimens, mostly in head and distal metaphysis...